Stu2p, the budding yeast member of the conserved Dis1/XMAP215 family of microtubule-associated proteins is a plus end–binding microtubule destabilizer

The Dis1/XMAP215 family of microtubule-associated proteins conserved from yeast to mammals is essential for cell division. XMAP215, the Xenopus member of this family, has been shown to stabilize microtubules in vitro, but other members of this family have not been biochemically characterized. Here we investigate the properties of the Saccharomyces cerevisiae homologue Stu2p in vitro. Surprisingly, Stu2p is a microtubule destabilizer that binds preferentially to microtubule plus ends. Quantitative analysis of microtubule dynamics suggests that Stu2p induces microtubule catastrophes by sterically interfering with tubulin addition to microtubule ends. These results reveal both a new biochemical activity for a Dis1/XMAP215 family member and a novel mechanism for microtubule destabilization

Stu2p binds tubulin and undergoes an open-to-closed conformational change

Stu2p from budding yeast belongs to the conserved Dis1/XMAP215 family of microtubule-associated proteins (MAPs). The common feature of proteins in this family is the presence of HEAT repeat–containing TOG domains near the NH2 terminus. We have investigated the functions of the two TOG domains of Stu2p in vivo and in vitro. Our data suggest that Stu2p regulates microtubule dynamics through two separate activities. First, Stu2p binds to a single free tubulin heterodimer through its first TOG domain. A large conformational transition in homodimeric Stu2p from an open structure to a closed one accompanies the capture of a single free tubulin heterodimer. Second, Stu2p has the capacity to associate directly with microtubule ends, at least in part, through its second TOG domain. These two properties lead to the stabilization of microtubules in vivo, perhaps by the loading of tubulin dimers at microtubule ends. We suggest that this mechanism of microtubule regulation is a conserved feature of the Dis1/XMAP215 family of MAPs

Disease-Associated Mutations in CEP120 Destabilize the Protein and Impair Ciliogenesis.

Ciliopathies are a group of genetic disorders caused by a failure to form functional cilia. Due to a lack of structural information, it is currently poorly understood how ciliopathic mutations affect protein functionality to give rise to the underlying disease. Using X-ray crystallography, we show that the ciliopathy-associated centriolar protein CEP120 contains three C2 domains. The point mutations V194A and A199P, which cause Joubert syndrome (JS) and Jeune asphyxiating thoracic dystrophy (JATD), respectively, both reduce the thermostability of the second C2 domain by targeting residues that point toward its hydrophobic core. Genome-engineered cells homozygous for these mutations have largely normal centriole numbers but show reduced CEP120 levels, compromised recruitment of distal centriole markers, and deficient cilia formation. Our results provide insight into the disease mechanism of two ciliopathic mutations in CEP120, identify putative binding partners of CEP120 C2B, and suggest a complex genotype-phenotype relation of the CEP120 ciliopathy alleles

Giant Lya nebulae associated with high redshift radio galaxies

We report deep Keck narrow-band Lya images of the luminous z > 3 radio galaxies 4C 41.17, 4C 60.07, and B2 0902+34. The images show giant, 100-200 kpc scale emission line nebulae, centered on these galaxies, which exhibit a wealth of morphological structure, including extended low surface brightness emission in the outer regions, radially directed filaments, cone-shaped structures and (indirect) evidence for extended Lya absorption. We discuss these features within a general scenario where the nebular gas cools gravitationally in large Cold Dark Matter (CDM) halos, forming stars and multiple stellar systems. Merging of these ``building'' blocks triggers large scale starbursts, forming the stellar bulges of massive radio galaxy hosts, and feeds super-massive black holes which produce the powerful radio jets and lobes. The radio sources, starburst superwinds and AGN radiation then disrupt the accretion process limiting galaxy and black hole growth, and imprint the observed filamentary and cone-shaped structures of the Lya nebulae.Comment: 36 Pages, including 8 Postscript figures. Accepted for publication in the Astrophysical Journa

Extended Lyα emission associated with 3C 294

We report the discovery of a large cloud of ionized gas associated with the radio galaxy 3C 294 at a redshift of 1.786. The radio source is a powerful double with a weak core. We detect Lyα emission with a total monochromatic luminosity of L_(Lyα) = 7.6 x 10^(44) ergs s^(-1) extended over ~100 x 170 kpc. The emission-line cloud is highly elongated and is well aligned with the inner radio source axis. Long-slit spectra, taken along the major axis, show spatially extended emission lines of N v λ1240, C IV λ1550, He II λ1640, and C III λ1909. The extended Lyα emission shows a large, smooth velocity gradient (1500 km s^(-1) and large intrinsic line widths (700-2600 km s^(-1)). The high-ionization lines, particularly C IV, show large velocity changes that are systematically different from those of Lyα. 3C 294 appears to be another, even more spectacular member of a class of powerful radio galaxies at high redshift typified by 3C 326.1. We discuss our observations from the point of view of star formation and the interaction of nuclear activity with ambient material. We consider two scenarios for the ionization of the Lyα cloud: ionization by a population of extremely massive stars and ionization by a non thermal continuum from the radio nucleus. Stellar photoionization appears to be insufficient in both the total number of ionizing photons and its ability to produce highly ionized species. We propose that the 150 kpc cloud of gas is photoionized by a central nonstellar source. We detect a very red compact object at 2.2 μm nearly coincident with the radio core. It is unclear whether the light from this object is primarily stellar or nonstellar

Metal-Enriched Gaseous Halos around Distant Radio Galaxies: Clues to Feedback in Galaxy Formation

We present the results of an optical and near-IR spectroscopic study of giant nebular emission-line halos associated with three z > 3 radio galaxies, 4C 41.17, 4C 60.07, and B2 0902+34. Previous deep narrowband Lyα imaging revealed complex morphologies with sizes up to 100 kpc, possibly connected to outflows and AGN feedback from the central regions. The outer regions of these halos show quiet kinematics with typical velocity dispersions of a few hundred km s^(-1) and velocity shears that can mostly be interpreted as being due to rotation. The inner regions show shocked cocoons of gas closely associated with the radio lobes. These display disturbed kinematics and have expansion velocities and/or velocity dispersions >1000 km s^(-1). The core region is chemically evolved, and we also find spectroscopic evidence for the ejection of enriched material in 4C 41.17 up to a distance of ≈60 kpc along the radio axis. The dynamical structures traced in the Lyα line are, in most cases, closely echoed in the carbon and oxygen lines. This shows that the Lyα line is produced in a highly clumped medium of small filling factor and can therefore be used as a tracer of the dynamics of high-redshift radio galaxies (HzRGs). We conclude that these HzRGs are undergoing a final jet-induced phase of star formation with ejection of most of their interstellar medium before becoming "red and dead" elliptical galaxies

Solution NMR assignment of the C-terminal domain of human chTOG

The microtubule regulatory protein colonic and hepatic tumor overexpressed gene (chTOG), also known as cytoskeleleton associated protein 5 (CKAP5) plays an important role in organizing the cytoskeleton and in particular in the assembly of k-fibres in mitosis. Recently, we dissected the hitherto poorly understood C-terminus of this protein by discovering two new domains—a cryptic TOG domain (TOG6) and a smaller, helical domain at the very C-terminus. It was shown that the C-terminal domain is important for the interaction with the TACC domain in TACC3 during the assembly of k-fibres in a ternary complex that also includes clathrin. Here we now present the solution NMR assignment of the chTOG C-terminal domain which confirms our earlier prediction that it is mainly made of α-helices. However, the appearance of the 1H–15N HSQC spectrum is indicative of the presence of a considerable amount of unstructured and possibly flexible portions of protein in the domain

The Spitzer High Redshift Radio Galaxy Survey

We present results from a comprehensive imaging survey of 70 radio galaxies at redshifts 1<z<5.2 using all three cameras onboard the Spitzer Space Telescope. The resulting spectral energy distributions unambiguously show a stellar population in 46 sources and hot dust emission associated with the active nucleus in 59. Using a new restframe S_3um/S_1.6um versus S_um/S_3um criterion, we identify 42 sources where the restframe 1.6um emission from the stellar population can be measured. For these radio galaxies, the median stellar mass is high, 2x10^11 M_sun, and remarkably constant within the range 13, there is tentative evidence for a factor of two decrease in stellar mass. This suggests that radio galaxies have assembled the bulk of their stellar mass by z~3, but confirmation by more detailed decomposition of stellar and AGN emission is needed. The restframe 500 MHz radio luminosities are only marginally correlated with stellar mass but are strongly correlated with the restframe 5um hot dust luminosity. This suggests that the radio galaxies have a large range of Eddington ratios. We also present new Very Large Array 4.86 and 8.46 GHz imaging of 14 radio galaxies and find that radio core dominance --- an indicator of jet orientation --- is strongly correlated with hot dust luminosity. While all of our targets were selected as narrow-lined, type 2 AGNs, this result can be understood in the context of orientation-dependent models if there is a continuous distribution of orientations from obscured type 2 to unobscured type 1 AGNs rather than a clear dichotomy. Finally, four radio galaxies have nearby (<6") companions whose mid-IR colors are suggestive of their being AGNs. This may indicate an association between radio galaxy activity and major mergers.Comment: 31 pages, 125 figures. Accepted for publication in the Astrophysical Journa

CCDC61/VFL3 Is a Paralog of SAS6 and Promotes Ciliary Functions.

Centrioles are cylindrical assemblies whose peripheral microtubule array displays a 9-fold rotational symmetry that is established by the scaffolding protein SAS6. Centriole symmetry can be broken by centriole-associated structures, such as the striated fibers in Chlamydomonas that are important for ciliary function. The conserved protein CCDC61/VFL3 is involved in this process, but its exact role is unclear. Here, we show that CCDC61 is a paralog of SAS6. Crystal structures of CCDC61 demonstrate that it contains two homodimerization interfaces that are similar to those found in SAS6, but result in the formation of linear filaments rather than rings. Furthermore, we show that CCDC61 binds microtubules and that residues involved in CCDC61 microtubule binding are important for ciliary function in Chlamydomonas. Together, our findings suggest that CCDC61 and SAS6 functionally diverged from a common ancestor while retaining the ability to scaffold the assembly of basal body-associated structures or centrioles, respectively